Pythiosis is a harmful disease caused by Pythium insidiosum, an aquatic oomycete. Therapeutic protocols based on antifungal drugs are often ineffective because the cytoplasmic membrane of P. insidiosum does not contain ergosterol. Therefore, the treatment of pythiosis is still challenging, particularly making use of natural products and secondary metabolites from bacteria. In this study, xanthyletin and substances obtained from Pseudomonas stutzeri ST1302 and Klebsiella pneumoniae ST2501 exhibited anti-P. insidiosum activity and, moreover, xanthyletin was non-toxic against human cell lines. The hyphae of P. insidiosum treated with these three substances exhibited lysis holes on a rough surface and release of anamorphic material. Therefore, xanthyletin could be considered a promising alternative agent for treating cutaneous pythiosis in the near future.

• MeSH
• Antifungal Agents pharmacology   MeSH
• Antiparasitic Agents pharmacology   MeSH
• Bacteria chemistry   metabolism   MeSH
• Cell Line MeSH
• Fibroblasts drug effects   MeSH
• Hyphae drug effects   ultrastructure   MeSH
• Complex Mixtures MeSH
• Coumarins pharmacology   MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Pythium drug effects   MeSH
• Cell Survival drug effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Pythium oligandrum is a soil born free living oomycete able to parasitize fungi and oomycetes prey, including important plant and animals pathogens. Pythium oligandrum can colonize endophytically the root tissues of diverse plants where it induces plant defenses. Here we report the first long-read genome sequencing of a P. oligandrum strain sequenced by PacBio technology. Sequencing of genomic DNA loaded onto six SMRT cells permitted the acquisition of 913,728 total reads resulting in 112X genome coverage. The assembly and polishing of the genome sequence yielded180 contigs (N50 = 1.3 Mb; L50 = 12). The size of the genome assembly is 41.9 Mb with a longest contig of 2.7 Mb and 15,007 predicted protein-coding genes among which 95.25% were supported by RNAseq data, thus constituting a new Pythium genome reference. This data will facilitate genomic comparisons of Pythium species that are commensal, beneficial or pathogenic on plant, or parasitic on fungi and oomycete to identify key genetic determinants underpinning their diverse lifestyles. In addition comparison with plant pathogenic or zoopathogenic species will illuminate genomic adaptations for pathogenesis toward widely diverse hosts.

• MeSH
• Beta vulgaris parasitology   MeSH
• Genome MeSH
• Proteome MeSH
• Pythium genetics   metabolism   MeSH
• Rhizosphere MeSH
• RNA-Seq MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Transcription factors (TFs) fine-tune the host defense transcriptome in response to pathogen invasions. No information is available on Zingiber zerumbet (Zz) TFs involved in defense response against Pythium myriotylum. Here, we provide a global identification, characterization, and temporal expression profiling of Zz TFs following an incompatible interaction with P. myriotylum using a transcriptome sequencing approach. We identified a total of 903 TFs belonging to 96 families based on their conserved domains. Evolutionary analysis clustered the Zz TFs according to their phylogenetic affinity, providing glimpses of their functional diversities. High throughput expression array analysis highlighted a complex interplay between activating and repressing transcription factors in fine-tuning Zz defense response against P. myriotylum. The high differential modulation of TFs involved in cell wall fortification, lignin biosynthesis, and SA/JA hormone crosstalk allows us to envisage that this mechanism plays a central role in restricting P. myriotylum proliferation in Zz. This study lays a solid foundation and provides valuable resources for the investigation of the evolutionary history and biological functions of Zz TF genes involved in defense response.

• MeSH
• Stress, Physiological MeSH
• Plant Immunity * MeSH
• Evolution, Molecular MeSH
• Pythium pathogenicity   MeSH
• Response Elements MeSH
• Plant Proteins genetics   metabolism   MeSH
• Transcription Factors genetics   metabolism   MeSH
• Transcriptome * MeSH
• Zingiberaceae genetics   immunology   microbiology   MeSH
• Publication type
• Journal Article MeSH

MAIN CONCLUSION: The level of resistance induced in different tomato genotypes after β-CRY treatment correlated with the upregulation of defence genes, but not sterol binding and involved ethylene and jasmonic acid signalling. Elicitins, a family of small proteins secreted by Phytophthora and Pythium spp., are the most well-known microbe-associated molecular patterns of oomycetes, a lineage of fungus-like organisms that include many economically significant crop pathogens. The responses of tomato plants to elicitin INF1 produced by Phytophthora infestans have been studied extensively. Here, we present studies on the responses of three tomato genotypes to β-cryptogein (β-CRY), a potent elicitin secreted by Phytophthora cryptogea that induces hypersensitive response (HR) cell death in tobacco plants and confers greater resistance to oomycete infection than acidic elicitins like INF1. We also studied β-CRY mutants impaired in sterol binding (Val84Phe) and interaction with the binding site on tobacco plasma membrane (Leu41Phe), because sterol binding was suggested to be important in INF1-induced resistance. Treatment with β-CRY or the Val84Phe mutant induced resistance to powdery mildew caused by the pathogen Pseudoidium neolycopersici, but not the HR cell death observed in tobacco and potato plants. The level of resistance induced in different tomato genotypes correlated with the upregulation of defence genes including defensins, β-1,3-glucanases, heveins, chitinases, osmotins, and PR1 proteins. Treatment with the Leu41Phe mutant did not induce this upregulation, suggesting similar elicitin recognition in tomato and tobacco. However, here β-CRY activated ethylene and jasmonic acid signalling, but not salicylic acid signalling, demonstrating that elicitins activate different downstream signalling processes in different plant species. This could potentially be exploited to enhance the resistance of Phytophthora-susceptible crops.

• MeSH
• Cyclopentanes metabolism   MeSH
• Ethylenes metabolism   MeSH
• Fungal Proteins metabolism   MeSH
• Host-Pathogen Interactions MeSH
• Salicylic Acid metabolism   MeSH
• Plant Leaves metabolism   microbiology   MeSH
• Plant Diseases microbiology   MeSH
• Oxylipins metabolism   MeSH
• Hydrogen Peroxide metabolism   MeSH
• Phytophthora MeSH
• Pythium MeSH
• Reactive Oxygen Species metabolism   MeSH
• Plant Growth Regulators metabolism   MeSH
• Signal Transduction * MeSH
• Solanum lycopersicum metabolism   microbiology   physiology   MeSH
• Publication type
• Journal Article MeSH

Pythium oligandrum (Oomycota) is known for its strong mycoparasitism against more than 50 fungal and oomycete species. However, the ability of this oomycete to suppress and kill the causal agents of dermatophytoses is yet to be studied. We provide a complex study of the interactions between P. oligandrum and dermatophytes representing all species dominating in the developed countries. We assessed its biocidal potential by performing growth tests, on both solid and liquid cultivation media and by conducting a pilot clinical study. In addition, we studied the molecular background of mycoparasitism using expression profiles of genes responsible for the attack on the side of P. oligandrum and the stress response on the side of Microsporum canis. We showed that dermatophytes are efficiently suppressed or killed by P. oligandrum in the artificial conditions of cultivations media between 48 and 72 h after first contact. Significant intra- and interspecies variability was noted. Of the 69 patients included in the acute regimen study, symptoms were completely eliminated in 79% of the patients suffering from foot odour, hyperhidrosis disappeared in 67% of cases, clinical signs of dermatomycoses could no longer be observed in 83% of patients, and 15% of persons were relieved of symptoms of onychomycosis. Our investigations provide clear evidence that the oomycete is able to recognize and kill dermatophytes using recognition mechanisms that resemble those described in oomycetes attacking fungi infecting plants, albeit with some notable differences.

• MeSH
• Antibiosis * MeSH
• Arthrodermataceae growth & development   MeSH
• Biological Therapy methods   MeSH
• Stress, Physiological MeSH
• Humans MeSH
• Microbial Viability MeSH
• Pilot Projects MeSH
• Pythium growth & development   MeSH
• Gene Expression Profiling MeSH
• Tinea therapy   MeSH
• Treatment Outcome MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Objectives Dermatophytosis, commonly known as ringworm, is a superficial fungal skin disease and zoonosis. Pythium oligandrum is a micromycete with mycoparasitic properties that is used in agriculture to control fungal infections on plants. Formulations containing P oligandrum were also developed for the treatment of dermatophytoses, but only a small number of case studies have been published. In order to document the process in simplified conditions in vitro, we investigated the effectiveness of P oligandrum against three pathogenic dermatophytes common in domestic animals. Methods Cultures of the pathogens grown on nutrient media and experimentally infected cat hair were treated with P oligandrum preparations in therapeutic concentration and the changes were documented by microscopic videos and scanning electron microscopy. Results There was strong mycoparasitic activity of P oligandrum against Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes. Conclusions and relevance P oligandrum was demonstrated to be effective against three common causes of dermatophytosis in vitro.

• MeSH
• Antifungal Agents pharmacology   MeSH
• Cats MeSH
• Microsporum drug effects   MeSH
• Cat Diseases drug therapy   microbiology   MeSH
• Parasitic Sensitivity Tests veterinary   MeSH
• Pythium * MeSH
• Tinea drug therapy   microbiology   veterinary   MeSH
• Trichophyton drug effects   MeSH
• Hair microbiology   MeSH
• Treatment Outcome MeSH
• Animals MeSH
• Check Tag
• Cats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

• Keywords
• Pythium oligandrum, mykoparazitismus,
• MeSH
• Antifungal Agents * adverse effects   therapeutic use   MeSH
• Biological Therapy veterinary   MeSH
• Biological Products MeSH
• Disinfection MeSH
• Pythium * MeSH
• Tinea * etiology   drug therapy   veterinary   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH

AIMS: Screening of bacterial flora for strains producing metabolites with inhibitory effects on the human pathogenic oomycete Pythium insidiosum. Separation and characterization of extracts from Pseudomonas stutzeri with anti-Pythium inhibitory activity. Search for genes with anti-Pythium effect within the genome of P. stutzeri. METHODS: A total of 88 bacterial strains were isolated from water resources in northeastern Thailand. Two screening methods were used to establish their inhibitory effects on P. insidiosum. One strain, P. stutzeri ST1302 was randomly chosen, and the extract with anti-P. insidiosum activity was fractionated and subfractionated using liquid column chromatography and purified by thin layer chromatography. The chemical structure of purified fractions was determined by Fourier transform infrared spectroscopy, nuclear magnetic resonance and mass spectrometry. Further, search for genes involved in the anti-Pythium activity (phenazine-1-carboxylic acid, 2,4-diacetylphloroglucinol, pyoluteorin and pyrrolnitrin) was undertaken in this P. stutzeri strain using primers described in the literature. RESULTS: Anti-P. insidiosum activity was detected in 16 isolates (18.2%). In P. stutzeri ST1302, a subfraction labeled PYK7 exhibited strong activity against this oomycete. It was assigned to the diketopiperazines as cyclo(D-Pro-L-Val). In the search for genes, one gene region was successfully amplified. This corresponded to pyrrolnitrin. The results suggest the possibility of using the related metabolites against P. insidiosum. This is the first report on the inhibitory effects of P. stutzeri against this oomycete. The results may contribute to the development of antimicrobial drugs/probiotics against pythiosis.

• MeSH
• Diketopiperazines pharmacology   MeSH
• Genome, Bacterial MeSH
• Microbial Sensitivity Tests MeSH
• Pseudomonas stutzeri chemistry   genetics   isolation & purification   MeSH
• Pyrrolnitrin pharmacology   MeSH
• Pythiosis drug therapy   microbiology   MeSH
• Pythium drug effects   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Thailand MeSH

• MeSH
• Bacterial Load MeSH
• Biofilms MeSH
• Lower Extremity MeSH
• Wound Healing * drug effects   MeSH
• Pharmaceutical Solutions therapeutic use   MeSH
• Humans MeSH
• Bandages, Hydrocolloid MeSH
• Pythium * MeSH
• Wounds and Injuries microbiology   MeSH
• Treatment Outcome MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Meeting Abstract MeSH

Tato kazuistika popisuje účinek léčby přípravkem Ecosin (registrovaná forma Pythium oligandrum pro zvířata) u kotěte britské kočky, u kterého byla klinicky a laboratorně diagnostikována dermatofytóza. Jako původce onemocnění bylo diagnostikováno Microsporum canis. Kotěti byla opakovaně aplikována koupel v přípravku Ecosin, který obsahuje živé kultury mykoparazitické houby Pythium oligandrum. Byly aplikovány tři celotělové koupele s týdenním odstupem a lokální aplikací přípravku, pokaždé druhý den po koupeli. Klinické vyšetření kotěte týden od ukončení aplikací již neprokázalo žádné příznaky dermatomykózy, negativní byly také kontrolní kultivace a trichoskopické vyšetření. Léčba přípravkem obsahujícím P. oligandrum byla bez jakýchkoli komplikací, bez nežádoucích i vedlejších účinků.

Presented case report describes therapeutical effect of biologic preparation Ecosin (registered form of mycoparasitic fungus Pythium oligandrum for animals) in a kitten of British shorthair cat, in which dermatophythosis has been diagnosed by means of clinical and laboratory examinations. Microsporum canis was identified as causal organism. The kitten was repeatedly treated by Ecosin in form of bath. Overall three baths were applied in a week interval and local administration of Ecosin was performed next day after each bath. No clinical symptoms were presented and both mycologic cultivation and trichoscopic examinations were negative after this medication. No adverse effect was recorded during and after treatment of the kitten by Ecosin.

• MeSH
• Dermatitis * diagnosis   therapy   veterinary   MeSH
• Clinical Laboratory Techniques MeSH
• Cats MeSH
• Microsporum * MeSH
• Mycoses * therapy   MeSH
• Cat Diseases MeSH
• Pythium * MeSH
• Zoonoses MeSH
• Animals MeSH
• Check Tag
• Cats MeSH
• Animals MeSH
• Publication type
• Case Reports MeSH